Kong Ling-Ling, Man Dong-Mei, Wang Tian, Zhang Guo-An, Cui Wen
Department of Pathology, Basic Science School, Jining Medical University, Jining, Shandong 272067, P.R. China.
Department of Gynecology, Affiliated Hospital of Jining Medical University, Jining, Shandong 272029, P.R. China.
Oncol Lett. 2016 Apr;11(4):2475-2480. doi: 10.3892/ol.2016.4244. Epub 2016 Feb 18.
The present study aimed to evaluate the effects of lysine-specific demethylase 1 (LSD1) downregulation, induced by small interfering RNA (siRNA) transfection, on the proliferation, colony formation, migration and invasion of the papillary thyroid carcinoma K1 cell line. The siRNA targeting LSD1 and scrambled non-targeting siRNA were each transfected into papillary thyroid carcinoma K1 cells. Downregulation of LSD1 mRNA and protein level was evaluated by reverse transcription-quantitative polymerase chain reaction, and immunocytochemical (ICC) analysis and western blotting, respectively. A Cell Counting kit-8 assay was applied to estimate the effect of LSD1-siRNA on cell growth. Migration and invasion abilities were estimated by Transwell chamber assay. A soft agar colony formation assay was performed to estimate the effect of LSD1-siRNA on tumorigenicity . ICC data showed that LSD1 protein was strongly expressed in the blank and control K1 cells compared with the LSD1-siRNA cells (F=15.192, P<0.01). Compared with the control cells, cells transfected with siRNA targeting LSD1 exhibited significant downregulation of LSD1 mRNA (t=6.845, P<0.01) and protein (F=53.764, P<0.01) levels. siRNA targeting LSD1 also downregulated cell proliferation following transfection for 24, 48 and 72 h (t=4.777, P<0.001; t=3.302, P=0.003; and t=3.017, P=0.006, respectively). Compared with the control group, the amount of cell invasion was gradually reduced in the LSD1-siRNA group (t=12.301, P<0.01). The number of migrating cells was significantly higher in the negative control group compared with the LSD1-siRNA group (t=7.911, P<0.01), and the ability of colony formation in the LSD1-siRNA cells was notably reduced in the soft agar formation assay (t=3.612, P=0.005). siRNA targeting LSD1 efficiently inhibits the proliferation, colony formation, migration and invasion of papillary thyroid carcinoma K1 cells.
本研究旨在评估通过小干扰RNA（siRNA）转染诱导赖氨酸特异性去甲基化酶1（LSD1）表达下调，对甲状腺乳头状癌K1细胞系的增殖、集落形成、迁移和侵袭的影响。将靶向LSD1的siRNA和无靶向作用的乱序siRNA分别转染至甲状腺乳头状癌K1细胞中。分别通过逆转录-定量聚合酶链反应、免疫细胞化学（ICC）分析和蛋白质印迹法评估LSD1 mRNA和蛋白水平的下调情况。应用细胞计数试剂盒-8法评估LSD1-siRNA对细胞生长的影响。通过Transwell小室试验评估迁移和侵袭能力。进行软琼脂集落形成试验以评估LSD1-siRNA对致瘤性的影响。ICC数据显示，与LSD1-siRNA细胞相比，LSD1蛋白在空白和对照K1细胞中高表达（F=15.192，P<0.01）。与对照细胞相比，转染靶向LSD1的siRNA的细胞LSD1 mRNA（t=6.845，P<0.01）和蛋白（F=53.764，P<0.01）水平显著下调。转染24、48和72小时后，靶向LSD1的siRNA也下调了细胞增殖（分别为t=4.777，P<0.001；t=3.302，P=0.003；t=3.017，P=0.006）。与对照组相比，LSD1-siRNA组细胞侵袭量逐渐减少（t=12.301，P<0.01）。与LSD1-siRNA组相比，阴性对照组迁移细胞数量显著更高（t=7.911，P<0.01），并且在软琼脂形成试验中，LSD1-siRNA细胞的集落形成能力显著降低（t=3.612，P=0.005）。靶向LSD1的siRNA可有效抑制甲状腺乳头状癌K1细胞的增殖、集落形成、迁移和侵袭。
